Recognition of endogenously synthesized HLA-DR4 restricted HCV epitopes presented by autologous EBV transformed B-lymphoblastoid cell line

• Published in: Vaccine 2005-01, Vol.23 (7), p.951-962
• Main Authors: Yang, Meiying, Zhu, Fenlu, Sønderstrup, Grete, Eckels, David D.
• Format: Article
• Language: English
• Subjects: Animals; Antigen Presentation - genetics; Antigens; Applied microbiology; B-Lymphocytes - immunology; B-Lymphocytes - metabolism; Base Sequence; Biological and medical sciences; Cell Line; Cell Line, Transformed; Endogenous antigen presentation; Epitopes, B-Lymphocyte - genetics; Epitopes, B-Lymphocyte - immunology; Epitopes, B-Lymphocyte - metabolism; Epstein-Barr virus; Fundamental and applied biological sciences. Psychology; HCV epitopes; Hepatitis; Hepatitis C Antigens - genetics; Hepatitis C Antigens - immunology; Hepatitis C Antigens - metabolism; Hepatitis C virus; Herpesvirus 4, Human - genetics; Herpesvirus 4, Human - immunology; HLA-DR4 Antigen - biosynthesis; HLA-DR4 Antigen - immunology; Humans; Mice; Mice, Transgenic; Microbiology; Minigene vaccine; Miscellaneous; Molecular Sequence Data; Peptides; T cell receptors; Vaccines; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects); Virology
• ISSN: 0264-410X; 1873-2518
• DOI: 10.1016/j.vaccine.2004.06.034

Hepatitis C virus (HCV) causes non-A, non-B hepatitis and infects an estimated 170 million people worldwide. The treatment for HCV infection is often unsuccessful with high costs and many side-effects. There is a great need for alternative therapies including preventive and therapeutic vaccination for HCV infection. The experiments in this study were carried out to elucidate whether endogenously expressed antigen can be presented to helper T-cells restricted by class II molecules and to determine whether responses to plasmid-derived antigen resemble those that we have reported for recombinant antigens or synthetic peptides. To address these issues, a multi-epitope minigene was expressed in 293T-cells and Epstein-Barr virus (EBV)-transformed B-lymphoblastoid cells (BLCL). The transfected BLCLs were employed as APCs to stimulate epitope-specific T-cell hybridomas (THC). The results demonstrated that the endogenously expressed minigene antigens could be processed and presented to T-cell hybridomas by HLA matched BLCL. Five out of seven incorporated epitopes were recognized. Blockade of HLA DR could abolish the release of IL-2, which demonstrated that the endogenously expressed minigene antigens were presented by MHC class II molecules. The presentation of endogenously expressed antigens was much more efficient than that of exogenous antigens, at least in the present study. The findings obtained here have important significance for the development of an HCV DNA vaccine.